CN114438190A - Opening and closing nerve soothing soup-autism core effect gene target and screening method thereof - Google Patents
Opening and closing nerve soothing soup-autism core effect gene target and screening method thereof Download PDFInfo
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Abstract
The invention discloses a key effect gene target of opening and closing tranquilization soup-autism and a screening method thereof, wherein an intersection target of an active ingredient and a disease is obtained through analysis of databases such as TCMSP (TCMSP) and uniprot, a protein-protein interaction (PPI) network is constructed, and a Cytoscape software is used for carrying out visual analysis on a result on an important module and a key target in the PPI network; carrying out gene GO and KEGG channel enrichment analysis on the target by adopting Metascape to obtain a visual graph of components, the target, gene functions, the channel and diseases. The invention solves the problems of large quantity of interaction between multiple active ingredients and multiple target proteins and difficult screening, and has important application significance for guiding the research and development of Chinese herbal compound.
Description
Technical Field
The invention relates to pharmacological research of a traditional Chinese medicine compound, in particular to a gene target for opening and closing nerve-soothing soup-autism core effect and a screening method thereof.
Background
Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder, frequently occurring before the age of 3 years, with individual phenotypes varying widely from mild to severe. In 2020, research reports issued by the centers for disease control and prevention (CDC) of the united states showed that the prevalence of autistic children was as high as 1:54 and showed a trend of rising year by year. However, there is still a lack of clinically definite diagnostic, prophylactic and therapeutic regimens for autism. Therefore, the pathogenesis of the autism is explored, a practical and effective treatment strategy is searched, a novel and effective treatment drug is developed, and the method has important research value and clinical significance for treating the autism.
The decoction for opening and closing tranquilization is a common prescription, and clinical data show that the decoction has obvious effect and can obviously improve the symptoms of autism children patients with 'heart and liver fire hyperactivity'. However, the specific active ingredients and pharmacological action mechanism of the start-stop and tranquilization decoction are not clear. The decoction mainly comprises polygala tenuifolia, rhizoma acori graminei, schisandra chinensis, coptis chinensis, lotus plumule and cinnamon, achieves the effects of clearing away heart-fire, calming the liver and tranquilizing the mind, and belongs to the field of treatment of heart-liver fire excess in traditional Chinese medicine.
Network pharmacology is an application tool which is developed in recent years and can be used for screening candidate genes or targets to research functions of related diseases and a drug treatment mechanism, and a novel drug development mode and a novel idea of a multi-drug and multi-target interaction relation are provided.
Disclosure of Invention
The invention aims to provide a gene target for opening and closing nerve-soothing soup-autism core action and a screening method thereof.
The technical scheme of the invention comprises the following steps:
the opening and closing nerve soothing soup-autism core effect gene target comprises the following genes:
VEGFA, IL6, TNF, INS, PTGS2, CXCL8, AKT1, MMP9, TP53 and EGF.
The application of the start-stop nerve-soothing soup-autism core effect gene target in preparation or screening treatment of autism.
The following active ingredients were screened as preferred subjects: quercetin, kaempferol, luteolin, 4' -methylhenbane Wuyama alkaloid, S- (2-carboxyethyl) -L-cysteine and oleic acid. .
A screening method for opening and closing nerve calming soup-autism core effect gene targets comprises the following operations:
1) screening active ingredients of the whole components of the mind-tranquilizing decoction;
2) screening action targets of the active ingredients of the start-stop nerve-soothing soup, and carrying out normalization and standardized naming on the targets to obtain potential targets of the active ingredients of the start-stop nerve-soothing soup;
3) screening an autism disease target in a disease target database;
taking the intersection gene of the potential target point of the active ingredients of the start-stop nerve-soothing soup and the target point of the autism disease as a candidate target point of the start-stop nerve-soothing soup for treating the autism;
4) constructing an active ingredient-disease target network for opening and closing the tranquilizing decoction: establishing a network diagram of the action target of the active ingredients of the decoction for opening and closing tranquilization corresponding to the candidate target of the decoction for opening and closing tranquilization for treating autism;
5) GO functional annotation and KEGG pathway enrichment analysis: performing gene function annotation and KEGG passage enrichment analysis on the intersection gene to determine the biological function related to the potential target;
constructing a target-path enrichment network based on the KEGG path enrichment result and the intersection target gene information, and prompting the start and stop of the tranquilization soup to correlate AGE-RAGE signal path, calcium signal path and dopaminergic nerve synapse with autism;
6) construction of protein-protein interaction PPI networks: uploading the intersection target genes to a String database, selecting various proteins, wisdom persons and confidence level options, obtaining protein-protein interaction PPI network data of the intersection target genes, and carrying out visual analysis on the result;
7) core gene and key compound screening and evaluation: the first 10 core genes are determined by an MCC method, and GO and KEGG channels are divided for the core genes.
The method specifically comprises the following operations:
1) according to a traditional Chinese medicine system pharmacological analysis database and a BATMAN-TCM database online platform, screening active ingredients for opening and closing the whole components of the tranquilizing decoction in the database under the condition that the bioavailability is more than or equal to 30% and the drug property index is more than or equal to 0.1;
2) screening action targets for opening and closing the active ingredients of the tranquilizing decoction by using a TCMSP online database, and carrying out normalization and standardized naming on the target data by using a UniProt database; 336 potential targets of the active ingredients of the six medicinal materials of the opening and closing nerve-soothing soup are obtained;
3) searching and screening a disease target gene as a candidate target of the disease in a disease target database;
the intersection gene of the potential target of the active ingredients of the start-stop nerve-soothing soup and the disease target is used as a candidate target of the start-stop nerve-soothing soup for treating autism;
4) constructing an active ingredient-disease target network for opening and closing the tranquilizing decoction: establishing a network diagram by corresponding the action target points of the active ingredients of the start-stop and tranquilization decoction to the candidate target points of the start-stop and tranquilization decoction for treating autism, analyzing and calculating the connection number by using a Cytoscape3.8.2 software system, and screening the first six ingredients according to the degree value;
5) GO functional annotation and KEGG pathway enrichment analysis: performing gene function annotation and KEGG channel enrichment analysis on the intersection gene through a metascape online database to determine the biological function related to the potential target; then, introducing the KEGG passage enrichment result and intersection target gene information into Cytoscape3.8.2 software, constructing a target-passage enrichment network, and realizing the visualization of the enrichment analysis result;
6) construction of protein-protein interaction PPI networks: uploading the obtained intersection target genes to a String database, selecting various proteins, wisdom and confidence 0.9 options, obtaining protein-protein interaction PPI network data of the intersection target genes, and performing visual analysis on the result by using cytoscape3.8.2 software;
7) core gene and key compound screening and evaluation: the first 10 core genes were determined by the MCC method in the "cytoHubba" plug-in of the Cytoscape3.8.2 software, and GO and KEGG pathway analysis was performed on the core genes by the metascape database.
The invention has the beneficial effects that:
the invention establishes a 'component-target-disease' system network diagram for treating autism by using active components of the opening and closing nerve-soothing soup for the first time based on a network pharmacological technology, and analyzes and determines how the opening and closing nerve-soothing soup plays a role through a 'multi-activity, multi-target and multi-channel' network system. The screening result shows that: core acting gene targets including VEGFA, IL6, TNF, INS, PTGS2, CXCL8, AKT1, MMP9, TP53, and EGF; the AGE-RAGE signal pathway, the calcium signal pathway and the dopaminergic synapse may be the main path for opening and closing the tranquilization decoction to treat autism.
The invention provides a new method and inspiration for the interaction of opening and closing the drug-target of the tranquilization decoction for treating autism, is beneficial to solving the problem that the interaction quantity between multiple active ingredients and multiple target proteins of a traditional Chinese medicine compound is too large, and the difficulty of one-by-one screening on experiments is difficult to achieve, and has important significance for guiding the research and development of the traditional Chinese medicine compound.
Drawings
FIG. 1 is a network diagram of open-close tranquilizing decoction-active compound-target spot-autism;
FIG. 2 is a schematic diagram of GO function annotation analysis of the intersection target gene of start-stop nerve-soothing soup and autism;
FIG. 3 is a schematic diagram of a KEGG pathway enrichment network for opening and closing the intersection target gene of the tranquilizing decoction and the autism;
FIG. 4 is a PPI network diagram of the intersection target gene of the on-off tranquilizing decoction and autism;
FIG. 5A is a diagram of identifying important blocks in a PPI network, and FIG. 5B is a diagram of a core gene network;
FIG. 6 is a network diagram of opening and closing tranquilizing decoction components, targets, pathways and diseases;
FIG. 7 is a flow chart of the study steps of start-stop tranquilizing decoction for treating autism.
Detailed Description
The present invention is further illustrated by the following specific examples.
The opening and closing nerve soothing soup-autism core effect gene target comprises the following genes:
VEGFA, IL6, TNF, INS, PTGS2, CXCL8, AKT1, MMP9, TP53 and EGF.
The application of the start-stop nerve-soothing soup-autism core effect gene target in preparation or screening treatment of autism.
The following active ingredients were screened as preferred subjects: quercetin, kaempferol, luteolin, 4' -methylhenbane Wuyama alkaloid, S- (2-carboxyethyl) -L-cysteine and oleic acid. .
A screening method for opening and closing nerve calming soup-autism core effect gene targets comprises the following operations:
1) screening active ingredients of the whole components of the mind-tranquilizing decoction;
2) screening action targets of the active ingredients of the start-stop nerve-soothing soup, and carrying out normalization and standardized naming on the targets to obtain potential targets of the active ingredients of the start-stop nerve-soothing soup;
3) screening an autism disease target in a disease target database;
taking the intersection gene of the potential target point of the active ingredients of the start-stop nerve-soothing soup and the target point of the autism disease as a candidate target point of the start-stop nerve-soothing soup for treating the autism;
4) constructing an active ingredient-disease target network for opening and closing the tranquilizing decoction: establishing a network diagram of the action target of the active ingredients of the decoction for opening and closing tranquilization corresponding to the candidate target of the decoction for opening and closing tranquilization for treating autism;
5) GO functional annotation and KEGG pathway enrichment analysis: performing gene function annotation and KEGG passage enrichment analysis on the intersection gene to determine the biological function related to the potential target;
constructing a target-path enrichment network based on the KEGG path enrichment result and the intersection target gene information, and prompting the start and stop of the tranquilization soup to correlate AGE-RAGE signal path, calcium signal path and dopaminergic nerve synapse with autism;
6) construction of protein-protein interaction PPI networks: uploading the intersection target genes to a String database, selecting various proteins, wisdom persons and confidence level options, obtaining protein-protein interaction PPI network data of the intersection target genes, and carrying out visual analysis on the result;
7) core gene and key compound screening and evaluation: the first 10 core genes are determined by an MCC method, and GO and KEGG channels are divided for the core genes.
The present invention is described in further detail below (the main steps are shown in the flow chart of FIG. 7)
1. Screening of active ingredients of start-stop nerve-soothing soup and prediction of action target thereof
1.1 screening method and analysis platform
Searching all chemical components of the six medicinal materials by using polygala tenuifolia, acorus gramineus, lotus plumule, schisandra chinensis, coptis chinensis and cinnamon as keywords according to a traditional Chinese medicine system pharmacological analysis platform (TCMSP), (https:// tcmspw.com/tcmsp.php) and BATMAN-TCM database, and establishing an open-close nerve calming soup chemical component database.
Oral Bioavailability (OB) refers to the relative amount of a drug reaching the systemic circulation after oral administration, and high oral bioavailability is an important indicator of whether a drug has biological activity; the drug-like property represents the similarity of a compound with known drugs.
The method takes the oral bioavailability threshold OB of more than or equal to 30 percent and the drug-like property of more than or equal to 0.1 as screening conditions, selects the compound with higher bioactivity by evaluating the in vivo process of the compound, and establishes an open-close nerve-soothing soup active ingredient database.
Potential action targets of the active ingredients are searched through a TCMSP database and a BATMAN-TCM database, target data are processed through a UniProt database (https:// www.uniprot.org /), and a species is selected as a person to be normalized and standardized.
The search of TCMSP database and BATMAN-TCM database shows that 45 active compounds meet the screening condition, wherein, the polygala root contains 6, the acorus gramineus contains 4, the lotus plumule contains 9, the schisandra (south) contains 5, the coptis contains 11, the cinnamon contains 10, some components are shared by 2 or more than 2 medicines, the shared components are removed, and the total amount of 44 active components is total. Specifically as described in table 1.
TABLE 1 effective active ingredients of decoction for tranquilizing mind
Searching potential action targets of 44 active ingredients through a TCMSP database and a BATMAN-TCM database, and carrying out normalization and standardized naming on target data through a UniProt database to finally obtain 336 potential targets of the active ingredients of the six medicinal materials; from the results, it can be seen that many compounds are predicted to be directed against the same protein target. The active compound and the potential target protein generate a network diagram, wherein the network diagram has 386 nodes and 978 edges.
2. Screening of autism disease targets
2.1 screening method and database platform
In three online databases of Gene Cards, Dispenet and OMIM, the Autism disease related target Gene is searched by taking 'Autism' as a keyword. GeneCards is a comprehensive database that automatically integrates data for approximately 125 network-derived genes (including genomic, transcriptomics, proteomics, etc.). Distenet is a discovery platform that contains one of the largest publicly available set of genes and variants associated with human disease. OMIM is an abbreviation for "0 nline Mendelian Inheritance in Man", namely, on-line "human Mendelian Inheritance", which is a database of human genes and genetic disorders. Finally, a common target shared by the three databases is retained as an autism related target, while other targets are deleted.
2.2 screening results and analysis
6790 autism disease-related target genes are screened out from three online databases, namely Gene Cards, Dispenet and OMIM. Combining the obtained target of the autism disease with the target of the compound active ingredient obtained in the item 1.1, and obtaining the active ingredient target-autism intersection target genes by using a Venny 2.1.0 online mapping tool, wherein the intersection target genes are regarded as potential targets of opening and closing tranquilization soup for treating autism. According to the opening and closing nerve calming soup target database and the autism related target database, 209 intersection genes are discovered together, and the genes are considered as potential action targets for opening and closing the nerve calming soup to treat autism.
3. Constructing an active compound-target-autism network for opening and closing the tranquilization decoction
3.1 visual analysis method for opening and closing nerve-soothing soup active compound-target-autism network
A cytoscape3.8.2 software is utilized to construct a compound-target-autism network for opening and closing the tranquilizing decoction, and the relation between an active compound and a potential target in the opening and closing the tranquilizing decoction is clarified.
In order to better understand the complex relationship between all bioactive compounds in the start-stop and tranquilization decoction and the related target points of autism, a network diagram of start-stop and tranquilization decoction active compound-target point-autism is established, as shown in fig. 1.
As shown in FIG. 1, it is found that 44 active ingredients in the start-stop tranquilizing decoction correspond to 209 potential targets according to network analysis, and 259 nodes are formed, and 742 interactions are performed. The number of connections was calculated using the analysis tool "NetworkAnalyzer" in Cytoscape 3.8.2. The first six ingredients are quercetin, kaempferol, luteolin, 4' -methylhenbane Wuyama alkaloid, S- (2-carboxyethyl) -L-cysteine and oleic acid.
4. GO functional annotation and KEGG pathway enrichment
4.1 analytical methods and databases
To investigate biological functions associated with potential targets, we performed Gene Ontology (GO) functional annotation and KEGG pathway enrichment analysis via the metascape online database (https:// metascape. org /). Metascape is a powerful gene function annotation analysis tool that can help us to apply the currently popular bioinformatics analysis method to the analysis of genes and proteins in batches. Among them, GO enrichment analysis was used to interpret and annotate genes by three-dimensional dimensions, including Cellular Component (CC), Molecular Function (MF), and Biological Process (BP) analysis. KEGG pathway enrichment is used primarily for pathway analysis. In addition, the species defined as "Human" contains at least three genes, and p values of 0.05 or less are considered to have statistical significance. And (4) reserving enrichment items with q values (namely corrected P values) less than 0.05, sorting the enrichment items according to the q values, and outputting a histogram. And (3) importing the KEGG passage enrichment result and intersection target gene information into Cytoscape3.8.2 software, constructing a target-passage enrichment network, and realizing the visualization of the enrichment analysis result.
In order to further verify whether the biological functions of the candidate targets are related to autism, GO and KEGG enrichment analysis is carried out through a metascape online database. Three GO entries were analyzed, including CC, MF, and BP, with the top 10 enrichment results for each entry shown in fig. 2. 2581 entries (P value < 0.05) were obtained by GO functional annotation analysis, including 2229 biological processes, 124 cell compositions, 228 molecular functions. The analysis finds that the biological process of the intersection target mainly relates to the reaction of cells to organic cyclic compounds, the reaction to injury, the reaction to inorganic substances, the circulatory system process, the inflammatory reaction and the like; cellular composition relates to the postsynaptic membrane, membrane rafts, receptor complexes, components of the postsynaptic membrane, and the like; molecular functions are mainly focused on neurotransmitter receptor activity, nuclear receptor activity, adrenergic receptor activity, catecholamine binding, and the like.
In addition, 347 KEGG pathways were found. The KEGG enrichment analysis shows that 209 potential targets are closely related to several key pathways related to autism, such as AGE-RAGE signaling pathway, calcium signaling pathway, CAMP signaling pathway, dopaminergic synapse, VEGF signaling pathway, cholinergic synapse, 5-hydroxytryptamine synapse, and the like. The first 20 abundant KEGG signal pathways are shown in figure 3. These findings suggest that calcium signaling pathways, CAMP signaling pathways, and 5-hydroxytryptamine ergic synapses may be the primary mechanism for opening and closing tranquilization soup for autism. And constructing a composition-target point-path-disease diagram of the opening and closing nerve-soothing soup according to the results for visual analysis, and referring to fig. 6.
5. Construction of protein-protein interaction (PPI) networks and screening of core targets
5.1 analysis method and database platform
Protein-protein interactions (PPI) are crucial in the regulation of biological systems, uploading the intersection target genes obtained in item 1.2 above to the String database. The result of selecting the option of Multiple proteins, setting the species as Homo sapiens and the confidence as 0.9, and selecting the hidden nodes to obtain the protein-protein interaction (PPI) network data of the intersection target genes is beneficial to understanding the mechanism related to the occurrence and development of diseases.
String is an online PPI analysis database that currently has the largest number of organisms (5090) and proteins (2460 million) and has a very broad and diverse baseline data source. Studies have shown that a small fraction of PPI networks with highly connected regions (subnetworks) have a higher probability of participating in bioregulation, while those nodes that are lightly connected do not play a critical role in the integrity of the entire network. Therefore, the result of the PPI network is imported into Cytoscape software for visual analysis, and all targets are clustered and analyzed by means of the MCODE plug-in unit to find important modules (subnets) in the network.
The MCODE selection criteria were:
cut-of value
＝2,node cut-of value
＝0.2,maximum depth＝100,and k-score＝2。
MCODE can detect densely connected regions in a large protein-protein interaction network that may represent molecular complexes.
In addition, further genetic confirmation that the core genes within the module are often significantly correlated with the disease state, and it is more meaningful to obtain the core genes from the PPI network than to screen individual genes without network information. Cytohubba ranks nodes in a network mainly through network functions, and provides 11-medium topology analysis methods including degree, maximum neighborhood components and the like, and MCC has better performance in the precision of predicting necessary protein from PPI network.
To further identify the central genes in potential targets, we identified the top 10 core genes using the MCC method in "cytoHubba" of the cytoscape3.8.2 software. Meanwhile, GO and KEGG path analysis is carried out on the core gene through a metascape database. Gene counts >3 and p values below 0.05 were considered as cutoff criteria. To reveal the relationship between the core gene and the possible therapeutic components.
The PPI network diagram of the intersection target gene of the open-close tranquilizing decoction and the autism is shown in figure 4, and 203 nodes and 2914 edges are shared. Two important modules with scores more than or equal to 5 are screened by MCODE, and key sub-networks and genes are screened according to the score ranking, wherein the higher the score is, the more key the genes are. See fig. 5A and 5B. And (3) performing calculation ranking through MCC functions in the cytoHubba plug-in, screening out core genes closely related to the autism, and determining the top ten core nodes.
The ten major core genes include VEGFA, IL6, TNF, INS, PTGS2, CXCL8, AKT1, MMP9, TP53, and EGF.
And then GO and KEGG enrichment analysis is carried out on the 209 intersection target genes of the start-stop nerve-soothing soup and the autism, so that the biological functions of the GO-stop nerve-soothing soup and the autism are verified, and the fact that the core gene is closely related to the autism is verified.
AKT1 participates in "calcium signal pathway", "neurotransmitter receptor activity" and "5-hydroxytryptamine synapsis", which are closely related to autism, six components of quercetin, kaempferol, luteolin, 4' -methylmercaptan, S- (2-carboxyethyl) -L-cysteine and oleic acid corresponding to central genes are identified in the decoction for opening and closing tranquilization, and quercetin from coptis chinensis and lotus plumule can be especially directed at most central genes.
Therefore, the application of the open-close tranquilizing decoction-autism core effect gene target in preparation or screening for treating autism is provided.
Further, the following action components were screened as preferable subjects: quercetin, kaempferol, luteolin, 4' -methylhenbane Wuyama alkaloid, S- (2-carboxyethyl) -L-cysteine and oleic acid.
The embodiments given above are preferred examples for implementing the present invention, and the present invention is not limited to the above-described embodiments. Any non-essential addition and replacement made by the technical characteristics of the technical scheme of the invention by a person skilled in the art belong to the protection scope of the invention.
Claims (7)
1. The opening and closing nerve calming soup-autism core effect gene target is characterized by comprising the following genes:
VEGFA, IL6, TNF, INS, PTGS2, CXCL8, AKT1, MMP9, TP53 and EGF.
2. The use of the open-close tranquilizing decoction-autism core action gene target of claim 1 in preparation or screening for treating autism.
3. Use according to claim 2, characterized in that the following active ingredients are screened as preferred subjects: quercetin, kaempferol, luteolin, 4' -methylhenbane Wuyama alkaloid, S- (2-carboxyethyl) -L-cysteine and oleic acid. .
4. A screening method for opening and closing nerve calming soup-autism core effect gene targets is characterized by comprising the following operations:
1) screening active ingredients of the whole components of the mind-tranquilizing decoction;
2) screening action targets of the active ingredients of the start-stop nerve-soothing soup, and carrying out normalization and standardized naming on the targets to obtain potential targets of the active ingredients of the start-stop nerve-soothing soup;
3) screening an autism disease target in a disease target database;
taking the intersection gene of the potential target point of the active ingredients of the start-stop nerve-soothing soup and the target point of the autism disease as a candidate target point of the start-stop nerve-soothing soup for treating the autism;
4) constructing an active ingredient-disease target network for opening and closing the tranquilizing decoction: establishing a network diagram of the action target of the active ingredients of the decoction for opening and closing tranquilization corresponding to the candidate target of the decoction for opening and closing tranquilization for treating autism;
5) GO functional annotation and KEGG pathway enrichment analysis: performing gene function annotation and KEGG passage enrichment analysis on the intersection gene to determine the biological function related to the potential target;
constructing a target-path enrichment network based on the KEGG path enrichment result and the intersection target gene information, and prompting the start and stop of the tranquilization soup to correlate AGE-RAGE signal path, calcium signal path and dopaminergic nerve synapse with autism;
6) construction of protein-protein interaction PPI network: uploading the intersection target genes to a String database, selecting various proteins, wisdom persons and confidence level options, obtaining protein-protein interaction PPI network data of the intersection target genes, and carrying out visual analysis on the result;
7) screening and evaluation of core genes and key compounds: the first 10 core genes are determined by an MCC method, and GO and KEGG channels are divided for the core genes.
5. The method for screening the open-close tranquilizing decoction-autism core action gene target according to claim 4, which specifically comprises the following operations:
1) according to a traditional Chinese medicine system pharmacological analysis database and a BATMAN-TCM database online platform, screening active ingredients for opening and closing the whole components of the tranquilizing decoction in the database under the condition that the bioavailability is more than or equal to 30% and the drug property index is more than or equal to 0.1;
2) screening action targets for opening and closing the active ingredients of the tranquilizing decoction by using a TCMSP online database, and carrying out normalization and standardized naming on the target data by using a UniProt database; 336 potential targets of the active ingredients of the six medicinal materials of the opening and closing nerve-soothing soup are obtained;
3) searching and screening a disease target gene as a candidate target of the disease in a disease target database;
the intersection gene of the potential target of the active ingredients of the start-stop nerve-soothing soup and the disease target is used as a candidate target of the start-stop nerve-soothing soup for treating autism;
4) constructing an active ingredient-disease target network for opening and closing the tranquilizing decoction: establishing a network diagram by corresponding the action target points of the active ingredients of the start-stop and tranquilization decoction to the candidate target points of the start-stop and tranquilization decoction for treating autism, analyzing and calculating the connection number by using a Cytoscape3.8.2 software system, and screening the first six ingredients according to the degree value;
5) GO functional annotation and KEGG pathway enrichment analysis: performing gene function annotation and KEGG channel enrichment analysis on the intersection gene through a metascape online database to determine the biological function related to the potential target; then, introducing the KEGG passage enrichment result and intersection target gene information into Cytoscape3.8.2 software, constructing a target-passage enrichment network, and realizing the visualization of the enrichment analysis result;
6) construction of protein-protein interaction PPI network: uploading the obtained intersection target genes to a String database, selecting various proteins, wisdom and confidence 0.9 options, obtaining protein-protein interaction PPI network data of the intersection target genes, and performing visual analysis on the result by using cytoscape3.8.2 software;
7) core gene and key compound screening and evaluation: the first 10 core genes were determined by the MCC method in the "cytoHubba" plug-in of the Cytoscape3.8.2 software, and GO and KEGG pathway analysis was performed on the core genes by the metascape database.
6. The method for screening the open-close tranquilizing decoction-autism core action gene target according to claim 5, wherein the first six components are as follows: quercetin, kaempferol, luteolin, 4' -methylhenbane Wuyama alkaloid, S- (2-carboxyethyl) -L-cysteine and oleic acid.
7. The method for screening the core action gene target of open-close tranquilizing decoction-autism according to claim 5, wherein the first 10 core genes comprise VEGFA, IL6, TNF, INS, PTGS2, CXCL8, AKT1, MMP9, TP53 and EGF.
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